EZ-Link™ NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns

Catalog number: 21450

Thermo Scientific™  Related applications: Protein Labeling & Crosslinking

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Description

Thermo Scientific EZ-Link NHS-PEG Solid Phase Biotinylation Kits provide an on-column approach to amine-targeted antibody biotinylation that provides greater control over the labeling reaction and clean-up process.

Features of the EZ-Link NHS-PEG Solid-Phase Biotinylation Kit:

Suitable for larger-scale reactions—1 to 10 mg antibody
Fast labeling and purification—the entire procedure takes only about one hour
Easy removal of spent and excess labeling reagent—simply wash away the reaction byproducts—no need for dialysis or gel filtration
No dilution effects—solid-phase method allows initially dilute antibodies to be recovered in a smaller volume after labeling
Optimized protocols—specific protocols for antibody ensure appropriate level of labeling (2 to 5 biotins per antibody molecule), minimizing possibility of inactivation caused by overlabeling
High-performance biotin reagent—NHS-PEG4-Biotin uses efficient, amine-specific NHS-ester chemistry and includes hydrophilic polyethylene glycol (PEG) spacer that stabilizes long-term solubility of labeled antibodies
Eight biotin-labeling experiments per kit—Thermo Scientific No-Weigh Packaging ensures that the biotin reagent is fully active for eight separate experiments

This innovative antibody-labeling system uses nickel-chelated agarose to temporarily immobilize antibody molecules via their histidine-rich Fc regions. Once held in place on the resin, the antibody can be biotinylated at primary amines with the high-quality, solubility-stabilizing NHS-PEG4-Biotin reagent. Excess labeling reagent and byproducts are then washed away before recovering the labeled and purified antibody from the resin using a mild imidazole solution. No gel filtration or dialysis is needed.

EZ-Link Solid-Phase Biotinylation Kits use an innovative strategy for antibody biotinylation that simplifies the process in many respects, making successful biotinylation possible for the novice and expert. The strategy involves immobilizing the antibody to a metal-chelated affinity support and performing the biotinylation in the solid phase.

Antibodies bind readily to metal-chelated supports, such as nickel or cobalt, binding primarily through the histidine-rich Fc region of IgG-class antibodies. This feature allows the derivatization chemistry to be performed easily while the antibody is bound to the support. This strategy facilitates reagent delivery and removal of spent reagent. The resulting modified antibody is then selectively removed from the support by elution with a buffered imidazole solution.

The solid-phase approach to antibody biotinylation provides more control over the reaction. For example: Reagent and antibody ratios and reaction times can be adjusted to achieve the proper degree of biotinylation for the intended application.

The kits contain an aqueous soluble amino group-specific reagent (NHS-PEG4-Biotin) that targets accessible amino (-NH2) groups on the surface of the antibody. The format can be applied at the multi-milligram antibody level in the 1 mL column kit (Part No. 21440) or to low microgram antibody levels for biotinylations using the mini-spin column kit (Part No. 21450).

Related Products
EZ-Link™ NHS-PEG Solid-Phase Biotinylation Kit - 1 mL Column
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Label or Dye: Biotin
Product Size: 8 reactions
Labeling Scale: 0.5-1 mg
Labeling Target: Antibodies (IgG)

Contents & storage

Sufficient For: 8 labeling reactions, each with 0.1 to 1 mg of antibody
• Ni-NTA Spin Columns, 0.2 mL, 8 columns
• PBS pack (makes 500 mL), 1 pack
• NHS-PEG4-Biotin, 8 x 2 mg
• Imidazole Stock Solution (4M), 5 mL
• Collection Tubes (2mL), 30 tubes

Store at 4°C.

Documents

Manuals & protocols